MA5-14505

antibody from Invitrogen Antibodies

Targeting: PGR NR3C3, PR

Provider product page for MA5-14505

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [140]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [2]
Immunohistochemistry [3]
Other assay [4]

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Product number: MA5-14505 - Provider product page
Provider: Invitrogen Antibodies
Product name: Progesterone Receptor Monoclonal Antibody (SP2)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-14505 targets Progesterone Receptor in IHC (P) and WB applications and shows reactivity with Human samples.
Antibody clone number: SP2
Concentration: 0.08 mg/mL

PGR protein structure - MA5-14505 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Progesterone Receptor was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1031637_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Progesterone Receptor was performed by loading 30 µg of T-47D Cas9 (Lane 1) and T-47D Progesterone Receptor KO(Lane 2) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Progesterone Receptor Monoclonal Antibody (SP2) (Product # MA5-14505, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution) using the iBright™ FL 1500 (Product # A44115). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Progesterone Receptor. Uncharacterized bands was observed in all the samples between ~ 45-80 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of T47D (lane 1), MCF 7 (lane 2) and MDA-MB-231 (lane 3). The blots were probed with Anti-Progesterone-Receptor Rabbit Monoclonal Antibody (Product # MA5-14505, 1:500-1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A 98 and 82 kDa band corresponding to Progesterone-Receptor was observed across the cell lines tested except in MDA-MB-231. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with PierceTM Power Blotter System (Product # 22834) The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Progesterone Receptor (green) in T47D cells. The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were stained with or without Progesterone Receptor rabbit monoclonal antibody (Product # MA5-14505), at a dilution of 1:50 for 1 hour at 37C, and then incubated with a Alexa Fluor 488 Superclonal goat anti-rabbit IgG secondary antibody (Product # A27034) at a dilution of 1:1000 for 30 minutes at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Progesterone Receptor was done on 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Progesterone Receptor (SP2) Rabbit Polyclonal Antibody (Product # MA5-14505) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human breast carcinoma stained with Progesterone Receptor using peroxidase-conjugate and DAB. Note strong nuclear staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Progesterone Receptor (SP2) showing staining in the nucleus of paraffin-embedded human uterus tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Progesterone Receptor Antibody (SP2) Rabbit Monoclonal Antibody (Product # MA5-14505) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Progesterone Receptor (SP2) showing staining in the nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Progesterone Receptor Antibody (SP2) Rabbit Monoclonal Antibody (Product # MA5-14505) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Immunohistochemical analysis. a Hematoxylin and eosin (H&E) staining of a cervical spinal schwannoma. Immunohistochemical staining for b estrogen alpha receptor; c estrogen beta receptor; d progesterone receptor; and e isotype (negative) controls. Scale bar: 50 mum

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 The expression of estradiol receptors and progesterone receptors in the uterus of rats after rhFSH treatment. For the detection of ( A ) estradiol receptor and ( B ) progesterone receptor by immunohistochemical staining. The degrees of staining were categorized as follows: (0), no staining; (1), faint staining; (2), moderate staining; and (3), strong staining. Arrows pinpointed the staining of ER or PR. Scale bar = 500 mum. G1: abdominal s.c. injection of normal saline twice daily for 4 days (days 1 to 4); G2: abdominal s.c. injection of rhFSH (1 IU) twice daily for 4 days (days 1 to 4); G3: vaginal s.c. injection of rhFSH (1 IU) twice daily for 4 days (days 1 to 4); G4: abdominal s.c. injection of rhFSH (4 IU) every other day (two injections) (days 1 & 3); G5: vaginal s.c. injection of rhFSH (4 IU) every other day (two injections) (days 1 & 3). Abbreviations: LE: luminal endometrium; GE: glandular endometrium; Myomet: myometrium.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7. Effects of puberty and steroid hormones on FOXA2 expression in Pgr Cre Foxa2cOE mice. (a) FOXA2 expression is lost from the LE of the uterus between PD 30 and 70. (b) Pgr Cre Foxa2cOE mice were ovariectomized on PD 30, rested for 10 days, and treated daily with subcutaneous injections of vehicle (sesame oil), P4 (1 mg), or E2 (100 ng) for 10 days (n = 4 mice per genotype per day). Immunoreactive FOXA2, PGR, Ki67, P63, and KRT14 were evaluated with immunohistochemistry.